Alveolar and lung liquid clearances were studied over 1, 4, and 6 h in intact anesthetized ventilated rats by instillation of 5% albumin solution with 1.5 microCi of 125I-labeled albumin (3 ml/kg into 1 lung or 6 ml/kg into both lungs). Alveolar protein clearance as measured by residual 125I-albumin in the lung over 6 h was similar to the slow rates measured in other species. Alveolar liquid clearance was estimated by the concentration of albumin in the air spaces. After 1 h, this concentration was 7.8 +/- 0.7 g/dl, which was significantly greater than the initial protein concentration of 5.3 +/- 0.2 g/dl (P < 0.05). Amiloride (10(-3) M) inhibited 45% of the basal alveolar liquid clearance, and ouabain (10(-3) M), instilled and intravenously infused (0.004 mg), inhibited 30% of the clearance. beta-Adrenergic agonist instillation increased alveolar liquid clearance to the fastest 1-h rate (48 +/- 3% of instilled volume) that we observed in any intact species. The removal of the instilled fluid from the lung (expressed as lung liquid clearance; 0.96 +/- 0.3 ml/h) was twice as fast as the rate of alveolar and lung liquid clearance reported in the isolated or in situ rat lung models. The rate of alveolar and lung liquid clearance in these intact rats was significantly faster than those in prior studies in dogs and sheep and was similar to the rates in rabbits.

The primary objective of these studies was to test the contribution of ventilation and blood flow to the removal of excess liquid from the air spaces and interstitium of the lung. First, after eliminating ventilation by clamping the left main bronchus in anesthetized sheep, alveolar and lung liquid clearance was not altered over 4 h compared with control sheep that were ventilated normally. Thus, removal of excess liquid across the alveolar epithelium was independent of the change in the transalveolar hydrostatic pressure gradient produced by ventilation. Second, to determine the effect of removing all blood flow to the lung, we developed a new in situ sheep lung model in which lung lymph flow was measured over 4 h with or without ventilation after the sheep had been exsanguinated. Alveolar liquid clearance, as measured by the percent increase in alveolar protein concentration over 4 h, was similar between sheep without blood flow (31 +/- 18%) compared with sheep with normal blood flow to the lungs (31 +/- 17%). Lung lymph flow contributed to only 10-15% of the clearance of the excess alveolar liquid that was transported to the interstitium, indicating that nonlymphatic pathways accounted for most of the excess lung liquid clearance in the absence of microvascular filtration. Third, because ouabain completely inhibited alveolar liquid clearance in this in situ sheep lung model, these data provide evidence that alveolar liquid clearance depends on an intact Na(+)-K(+)-ATPase-dependent pump mechanisms. Finally, this in situ model represents a unique experimental preparation that can be used to study the alveolar epithelial barrier without blood flow or ventilation for a short time (4 h) interval.

IntroductionThe overall objective of these studies was to determine whether IgG antibody to Pseudomonas aeruginosa would modify the acute lung and pleural injury that developed over 24 h after the instillation of 1010 live P. aeruginosa into the distal airspaces of one lung in unanesthetized sheep. Using a quantitative experimental model to measure protein permeability across the alveolar epithelial, lung endothelial, and pleural mesothelial barriers, the effect of IgG antibody to P. aeruginosa was examined under four different experimental conditions. First, the effect of IgG antibody to P. aeruginosa in the circulation was examined by instilling 1010 live P. aeruginosa in 5% ovine albumin in sheep that had been vaccinated. Under these conditions, the presence of circulating IgG antibody to P. aeruginosa reduced lung endothelial injury but did not modify the lung epithelial or pleural injury caused by intraalveolar P. aeruginosa. Therefore, the second experimental protocol determined the effect of instilling immune serum from a sheep that had been vaccinated so that IgG antibody to P. aeruginosa was present in both the circulation and in the airspaces along with instillation of live bacteria. Under these conditions, injury to the lung endothelium, alveolar epithelium, and pleural space was completely prevented. Therefore, the third protocol examined the protective effect of instillation of IgG antibody to P. aeruginosa in the airspaces concurrent with the live bacteria. Interestingly, intraalveolar IgG antibody to P. aeruginosa prevented all evidence of lung epithelial and pleural injury, and this effect was associated with a marked decrease in the number of viable bacteria in the lung after 24 h. Therefore, the fourth protocol examined the prophylactic effect of instillation of the specific IgG antibody to P. aeruginosa 24 h before instillation of the bacteria. With this prophylactic regimen, epithelial, endothelial, and pleural injury were prevented, and there was a significant decrease in the number of bacteria recovered from the lung. (3)(4)(5). These studies have also suggested that the most protective antibodies against P. aeruginosa are opsonic rather than bactericidal, thus providing a rationale for P. aeruginosa immunization strategies that emphasize augmentation of humoral rather than cellmediated immunity (3). In contrast to older vaccines, which contained the whole endotoxin antigen, a Pseudomonas vaccine containing a high molecular weight, immunogenic 0-specific polysaccharide antigen, but lacking the toxic lipid A component, has been shown to be well tolerated and immunogenic in normal human volunteers (6). This vaccine also has been shown to achieve a plasma level of IgG antibody to P. aeruginosa comparable to the levels measured in patients who survived P. aeruginosa sepsis (7,8). Also, immunization with P. aeruginosa 0-specific polysaccharide antigen has improved survival after a homologous bacterial challenge in both burned and granulocytopenic mice and in a model of pneumonia in gui...

The alveolar epithelium is not injured by the apical application of moderate doses of Pseudomonas aeruginosa strains that produce protease. To determine the effect of Pseudomonas proteases on the basolateral surface of the alveolar epithelium, a series of experiments were done, in which P. aeruginosa strains that produce and do not produce proteases were administered intravenously. Subsequently, an innocuous dose of bacteria was instilled into the lungs of the rabbits. Although all the intravenous Pseudomonas strains increased the extravascular lung water to a similar degree, only the intravenous administration of the protease-producing P. aeruginosa strain increased the vulnerability of the alveolar epithelium to injury by the subsequent airspace bacteria. Bacteremia secondary to P. aeruginosa strains producing proteases could increase the chances of developing acute lung injury.